PřF:Bi5599c App. Biochem. and Cell Meth pr - Course Information

Bi5599c Applied Biochemistry and Cell Biology Methods - practice

Extent and Intensity

0/2/0. 2 credit(s) (plus extra credits for completion). Type of Completion: z (credit).

Teacher(s)

Mgr. Jiřina Medalová, Ph.D. (seminar tutor)
RNDr. Vendula Hlaváčková Pospíchalová, Ph.D. (seminar tutor)
RNDr. Petra Křížková (seminar tutor)

Guaranteed by

prof. Mgr. Vítězslav Bryja, Ph.D.
Department of Experimental Biology – Biology Section – Faculty of Science
Contact Person: Mgr. Jiřina Medalová, Ph.D.
Supplier department: Department of Experimental Biology – Biology Section – Faculty of Science

Timetable

Mon 14:00–15:50 D36/215

Prerequisites

Bi3030 Animal physiology && NOW ( Bi5599 App. Biochem. and Cell Methods )
The subject follows and belongs to the logical unit of practical courses of Animal Physiology and Special Methods of Animal Physiology.

Course Enrolment Limitations

The course is only offered to the students of the study fields the course is directly associated with.

fields of study / plans the course is directly associated with

there are 12 fields of study the course is directly associated with, display

Course objectives

The aim of this practicals is to teach students to: perform both the classical and modern techniques used in basic and applied research (see Syllabus); use up-to-date trends and modern techniques used in basic research of molecules, specific cells and cell populations; evolve their laboratory skills; decide what is the most appropriate assay with newly obtained methodological background; work in well-equipped facilities; use obtained knowledge and skills in experiments designed for their bachelor and diploma thesis

Learning outcomes

At the end of the course students should be able to: perform both the classical and modern techniques used in basic and applied research (see Syllabus); use up-to-date trends and modern techniques used in basic research of molecules, specific cells and cell populations; evolve their laboratory skills; decide what is the most appropriate assay with newly obtained methodological background; work in well-equipped facilities; use obtained knowledge and skills in experiments designed for their bachelor and diploma thesis

Syllabus

• 1. Cultivation and use of cell cultures a) Lab work safety, how to use flow box, cell splitting, seeding and treatment - modification of beta catenin activity b) Transfection of cells by PEI polycation - assessment of transfection efficiency, TopFlash method for assessment of beta catenin activity 2. Gene expression evaluation by PCR a) Isolation of RNA by affinity purification, assessment of concentration and purity and stability of isolated RNA b) Reverse transcription of mRNA to cDNA c) Quantitative real time PCR for expression of beta catenin target genes, confirmation of reaction specificity by melting curve, methods for evaluation of results, counting of solution concentrations, cell suspensions dilution etc. 3. Assessment of protein expression by Western blotting a) Preparation and quantification of SDS lysates b) Separation of protein fragments by SDS PAGE c) Wet blotting of proteins to PVDF membrane d) Immunodetection of proteins - beta catenin and receptors involved to its regulation

Literature

• Cell culture and upstream processing. Edited by M. Butler. 1st ed. New York: Taylor & Francis, 2007, xiv, 187. ISBN 9780415399692. info
• Mammalian cell culture : essential techniques. Edited by A. Doyle - J. B. Griffiths. Chichester: John Wiley & Sons, 1997, xv, 173. ISBN 0471970573. info
• Cancer cell culture : methods and protocols. Edited by Simon P. Langdon. Totowa, N.J.: Humana Press, 2004, xii, 360. ISBN 1588290794. info
• Animal cell culture :a practical approach. Edited by R. Ian Freshney. 2nd ed. Oxford: Oxford University Press, 1992, xix, 329 s. ISBN 0-19-963213-8. info
• Cell biological appplications of confocal microscopy. Edited by Brian Matsumoto. 2nd ed. Amsterdam: Academic Press, 2002, xii, 507. ISBN 0125804458. info
• Molecular cloning, A laboratory manual, Vol. 1-3, Eds. J. Sambrook and D. W. Russell, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York 2001
• Short protocols in molecular biology, Vol. 1 and 2, 5th edition, John Wiley and Sons, Inc., 2002
• Current protocols in cytometry, Vol. 1 and 2, Ed. J. P. Robinson, John Wiley and Sons, Inc., 1997
• Methods in Cell Biology, Vol. 75, Cytometry, 4th edition, New developments, Eds. Z. Darzynkiewicz, M. Roederer, H. J. Tanke, Elsevier Academic Press, 2004
• CROWTHER, John R. The ELISA guidebook. Totowa, N.J.: Humana Press, 2001, xi, 421. ISBN 0896037282. info

Teaching methods

Short theoretic lectures followed by practical course of selected specific methods; demonstrative introduction to advanced methodologies; homeworks; class discussion with skilled teacher.

Assessment methods

During the course students must complete given protocols with filled results and answers.

Language of instruction

Czech

Follow-Up Courses

• Bi5611c Special methods of animal physiology

Further comments (probably available only in Czech)

Study Materials
The course is taught annually.
Information on course enrolment limitations: Na předmět se vztahuje povinnost registrace, bez registrace může být znemožněn zápis předmětu!

• Enrolment Statistics (Autumn 2019, recent)
  
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